F BCL1; FP BCL1
t(11;14)(q13;q32), IGH/CCND1; Cyclin D1
B-cell lymphoma, Non-Hodgkin lymphoma (NHL), Mantle cell lymphoma (MCL)
Fluorescence in situ Hybridization (FISH)
3-5 days
F BCL1
5mL peripheral blood in sodium heparin
3mL bone marrow in sodium heparin
Fixed cytogenetically prepared cells in sterile centrifuge tube with pellet visible in 3:1, Methanol:Acetic Acid
FP BCL1
FFPE tissue is acceptable for FISH analysis. Preferred fixative is 10% neutral buffered formalin. Tissues preserved in B5 fixative or decalcified are usually not suitable for FISH. Tumor sections cut 3-5 µm thick and mounted on positively charged organosilane coated (silanized) slides work well. Request several unstained sections (two for each probe) and one H&E stained slide
4°C to 25°C during transit, but specimens may be transported on refrigerated gel packs. Do not allow the gel pack to come in contact with the specimen. Do not freeze. Extreme temperatures should be avoided.
Clotted specimen; Specimen exposed to extreme temperature; Anticoagulant toxic to cells; Insufficient number of cells; Improper fixative
FISH results indicate whether translocation is present or absent. A normal signal pattern results in two orange signals and two green signals. A t(11;14) results in a signal pattern of two orange/green (yellow) fusions, one on each of the abnormal chromosomes 11 and 14 and single orange and green signals from the normal chromosomes.
Mantle cell lymphoma can be difficult to distinguish morphologically from other B-cell malignant lymphomas. The translocation probe is designed to detect the juxtaposition of the IGH locus (14q32) next to the CCND1 (cyclin D1) gene (11q13) in both metaphase and interphase nuclei. This altered location of the IGH locus is thought to cause over expression of cyclin D1 in mantle cell lymphomas.
The t(11;14)(q13;q32) can be detected in approximately 95% of mantle cell lymphomas by FISH analysis. In contrast, PCR can detect the t(11;14) in 30-60% of cases, chromosome analysis can detect up to 70%, and Southern blot up to 50% of the translocations. This assay is an excellent alternative to immunoperoxidase detection of cyclin D1 over expression.